Proteomic and Functional Analyses of Astrocyte Exosomes Astrocytes are the most abundant cell type in the central nervous system (CNS). Over the past decade increasing attention has been paid to the critical homeostatic and diverse roles astrocytes play in the CNS health and disease. The emerging influence of astrocytes in CNS physiology and pathology has been highlighted by how factors secreted by astrocytes impact CNS development, synaptic transmission, neurodegeneration, or infection. Hence, understanding how astrocytes respond to and are regulated by extracellular cues is expected to identify fundamental mechanisms that govern how the CNS adapts and responds to injury or inflammation. Exosomes are a class of very small extracellular vesicles that are known to be secreted by many cell types. Recent studies have now identified critical functions for exosomes in mediating cell signaling and inflammatory responses. However, little is known about the function of exosomes released from astrocytes in response to inflammation and no previous study has detailed what proteins are contained in astrocytic exosomes. This gap in our knowledge will be addressed in this application as follows:
Specific Aim 1 will generate a proteomic map of the proteins contained within exosomes isolated from astrocytes, under basal and inflammatory conditions, using a cutting edge proteomic fractionation platform.
Specific Aim 2 will determine the function of astrocyte exosomes as putative mediators of astrocytic responses to inflammatory stimuli both in vitro and in vivo. We provide preliminary data that demonstrate a qualitative and quantitative increase in the release of exosomes from cultured astrocytes in response to treatment with interleukin-1beta. Importantly, we present data to indicate that astrocyte-derived exosomes are present within systemic circulation and can be collected and quantified from blood serum. We hypothesize that exosomes represent a fundamental component of the astrocytic response to inflammatory and neuropathological conditions. Results from these studies will provide a new functional basis to understand how exosomes define astrocyte function. This work will provide the scientific community with two shared outcomes: (1) we will produce a proteomic map of astrocytic exosomes under basal and inflammatory conditions that might lead to high fidelity biomarker identification, and (2) we will determine the functional role of astrocytic exosomes in in vitro and in vivo models. Thus, these studies may stimulate widespread interest in astrocyte-derived exosomes as mediators of intercellular communication in the CNS in health and disease.
Exosomes are small extracellular vesicles released by cells as a form of intercellular communication that can fuse with other cells to influence their function Astrocytes are the most plentiful cells in the nervous system and astrocyte-derived exosomes can be found in blood. Since we do not know the contents or function of these astrocyte exosomes, this project will use cutting edge research tools to develop our understanding of astrocyte exosomes by testing the role of astrocyte-derived exosomes on cell-to-cell signaling and inflammatory damage in the nervous system.
|Nicaise, Alexandra M; Johnson, Kasey M; Willis, Cory M et al. (2018) TIMP-1 Promotes Oligodendrocyte Differentiation Through Receptor-Mediated Signaling. Mol Neurobiol :|
|Nicaise, Alexandra M; Bongarzone, Ernesto R; Crocker, Stephen J (2016) A microglial hypothesis of globoid cell leukodystrophy pathology. J Neurosci Res 94:1049-61|
|Johnson, Kasey M; Crocker, Stephen J (2015) TIMP-1 couples RhoK activation to IL-1?-induced astrocyte responses. Neurosci Lett 609:165-70|
|Johnson, Kasey M; Milner, Richard; Crocker, Stephen J (2015) Extracellular matrix composition determines astrocyte responses to mechanical and inflammatory stimuli. Neurosci Lett 600:104-9|